Jackie, while no longer in vogue (due to the popularity/ease of GFP), beta-gal remains an excellent choice for reporter gene detection. It is considerably more sensitive than GFP; we were able to detect as few as 5 molecules of beta-gal per cell: probably a single mRNA molecule's worth! However, the beta-gal assay requires just a little bit more work than GFP. A full description of the methodology can be found in a few references. Also see the chapter in Current Protocols for Cytometry, which updates a review of these articles plus a variety of articles that have used the method in the past 10 years. 1. Nolan, G. P., et al. (1988). Fluorescence-activated cell analysis and sorting of viable mammalian cells based on beta-D-galactosidase activity after transduction of Escherichia coli lacZ. Proc Natl Acad Sci U S A 85:2603. 2. Fiering, S., et al. (1991). Improved FACS-Gal: flow cytometric analysis and sorting of viable eukaryotic cells expressing reporter gene constructs. Cytometry 12:291. 3. Roederer, M., et al. (1991). FACS-Gal: Flow cytometric analysis and sorting of cells expressing reporter gene constructs. Methods: A Companion to Methods in Enzymology 2:248. mr
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